Structural insights into pandemic and emerging influenza viruses

对大流行和新出现的流感病毒的结构见解

基本信息

批准号：
8644586
负责人：
IAN A WILSON
金额：
$ 82.26万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-04-15 至 2015-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8644586
关键词：
Affect Amino Acids Animals Antibodies Binding Biochemical Birds Cells Centers for Disease Control and Prevention (U.S.)Chiroptera Collaborations Complement Complex Crystallography DNA-Directed RNA Polymerase Development Disease Progression Economic Burden Electron Microscopy Epidemic Epitopes Equilibrium Event Evolution Exhibits Family suidae Future Genetic Polymorphism Genetic Transcription H7N7 Head Hemagglutinin Human Immune response Immune system Infection Influenza Influenza A Virus, H1N1 Subtype Influenza A Virus, H5N1 Subtype Influenza A virus Investigation Knowledge Laboratories Lead Mediating Membrane Glycoproteins Methodology Molecular Mutation Neuraminidase Nucleoproteins Pathogenesis Pathogenicity Phylogenetic Analysis Polymerase Polysaccharides Population RNA RNA chemical synthesis Reagent Research Ribonucleoproteins Ruthenium Ben Serotyping Site Specificity Structure Substrate Specificity Surface Antigens Technology Testing Therapeutic Intervention Tropism Vaccination Vaccines Viral Viral Genome Virulence Virulence Factors Virus Virus Diseases Virus Receptors base combat combinatorial design flu immunogenicity influenzavirus inhibitor/antagonist insight neutralizing antibody new technology novel novel strategies novel therapeutics novel vaccines pandemic disease pandemic influenza prevent receptor receptor binding response scaffold seasonal influenza small molecule stem swine flu vaccine candidate virus development

项目摘要

DESCRIPTION (provided by applicant): Influenza A viruses exhibit extreme diversity via multiple serotypes of the hemagglutinin (HA 1-16) and neuraminidase (NA 1-9) surface antigens. To date, only three of the possible 144 combinations found in bird and animal reservoirs have been associated with human pandemics (H1N1, H2N2, H3N2). Recently, a distinct lineage of influenza A viruses has been identified in bats, further increasing the spectrum of possible zoonotic viruses that could infect humans. This proposal seeks to elucidate at the structural level, key sites of vulnerability on influenza virus for development ofa sustainable cross-serotype immune response, and understand activity relationships of the surface glycoproteins (HA, NA) and ribonucleoprotein (RNP) complex, including the polymerase (PA, PB1, PB2), that underlie the pathogenicity and transmissibility of pandemic and seasonal influenza viruses. Antibody-mediated neutralization of influenza virus is a complex combinatorial problem for the human immune system as it is presented with diverse, highly variable and constantly evolving viruses. While neutralizing antibodies against human flu are traditionally regarded as being strain specific, recent studies have shown that a much broader response can be mounted over decades of evolution of a particular subtype (e.g. H3N2), across group 1 or group 2, and even across two major phylogenetic groups (I and 2). While these examples provide compelling evidence that the immune system is capable of mounting a sustained, cross-serotype response against influenza, how to elicit broadly neutralizing antibodies by vaccination is poorly understood. Therefore, we propose to determine the structural basis of broad neutralization and delineate the sites of vulnerability on the HA to enable development of novel vaccine scaffolds and even small molecule inhibitors that ameliorate or prevent disease progression. Furthermore, because we do not understand why certain viruses, such as the recent H1N1 2009 swine flu, are able to enter the human population and cause pandemics, we will also study the molecular basis of pathogenicity. Using a novel strategy to investigate both HA and NA substrate specificity and activity using newly designed glycan microarrays, we will assess the functional relationships between NA and HA activity. In this way, we will test the hypothesis that efficient infection of humans by influenza viruses requires a functional balance between the binding and specificity of HA and enzymatic activity of the NA with host glycan receptors. Another key factor in host-specific pathogenicity is the replication machinery composed of the RNP with associated polymerase, where mutations can alter polymerase activity and interaction with host cell factors. Structural and functional understanding of the RNP will enable other approaches to combat influenza infection. A combined biophysical and biochemical approach from three laboratories employing state of the art x-ray crystallography, electron microscopy and glycan array technologies will be used to provide key insights into influenza virus neutralization, tropism and pathogenesis, to reveal novel strategies to control and combat future pandemics.

描述（由申请人提供）：流感病毒通过血清凝集素（HA 1-16）和神经氨酸酶（Na 1-9）表面抗原表现出极端多样性。迄今为止，在鸟类和动物储层中发现的144种组合中只有三种与人类大流行有关（H1N1，H2N2，H3N2）。最近，在蝙蝠中鉴定出了流感A病毒的独特谱系，进一步增加了可能感染人类的人畜共患病毒的谱系。 This proposal seeks to elucidate at the structural level, key sites of vulnerability on influenza virus for development ofa sustainable cross-serotype immune response, and understand activity relationships of the surface glycoproteins (HA, NA) and ribonucleoprotein (RNP) complex, including the polymerase (PA, PB1, PB2), that underlie the pathogenicity and transmissibility of pandemic and seasonal流感病毒。抗体介导的流感病毒中和是人类免疫系统的复杂组合问题，因为它具有多种，高度可变且不断发展的病毒。尽管传统上认为对人流感的中和抗体被视为特定于菌株，但最近的研究表明，在数十年的特定亚型（例如，H3N2），第1组或第2组，甚至在两个主要的系统发生组（I和2）中，可以安装更广泛的反应（例如H3N2）。尽管这些例子提供了令人信服的证据，表明免疫系统能够安装针对流感的持续的，跨色谱的反应，但如何通过疫苗接种而广泛地中和抗体。因此，我们建议确定广泛中和的结构基础，并描绘出HA上脆弱性的部位，以便能够发展出新的疫苗支架，甚至可以改善或防止疾病进展的小分子抑制剂。此外，由于我们不明白某些病毒（例如最近的H1N1 2009猪流感）能够进入人群并引起大流行病，因此我们还将研究致病性的分子基础。使用新型策略，使用新设计的聚糖微阵列研究HA和NA底物的特异性和活性，我们将评估Na和HA活性之间的功能关系。这样，我们将检验以下假设：流感病毒对人类的有效感染需要在HA的结合和特异性与NA与宿主聚糖受体的酶活性之间保持功能平衡。宿主特异性致病性的另一个关键因素是由RNP与相关聚合酶组成的复制机制，其中突变可以改变聚合酶活性并与宿主细胞因子相互作用。对RNP的结构和功能理解将使其他方法能够打击流感感染。三个实验室采用了X射线晶体学，电子显微镜和聚糖阵列技术的三个实验室的合并生物物理和生化方法，可用于提供对流感病毒中和，热带主义和发病机理的关键见解，以揭示新的策略来控制和梳理未来的Pandemics。